DESC:FIELD OF INVENTION
The current invention generally relates to ready to eat food, and in particular to a millet-based flatbread and preparation method thereof.

BACKGROUND
Rapid pace of urbanization compounded with the rise in small or nuclear families has accelerated the growth of ready to eat (RTE) food market. The growing demand for convenience in food among consumers who are hard-pressed for time is a key driver for the RTE food market.
RTE food typically contains preservatives and other chemicals for retaining the freshness of food. The preservatives are chemicals added to increase shelf-life of a food product. With increase in consumer interest for food products containing minimal chemicals, there is renewed efforts in food industry to meet this requirement.
Flatbreads in one or the other form is consumed all around the world, such as tortillas, naans, rotis and the like. The flatbread-based RTE food are often sold frozen, which can be consumed at a later time after warming, or in some instances after cooking. The RTE flatbreads which are sold at room temperature are often loaded with preservatives to prolong their shelf life from 1 to 2 days to at most a week. Moreover, the RTE flatbreads lack the taste and texture of freshly made ones.
Minor cereals, such as millets, are important crops in semiarid and tropical regions of the world due to their resistance to pests and diseases, short growing season, and productivity under hardy and drought conditions where major cereals cannot be
relied upon to provide sustainable yields. However, millets have remained largely neglected in commercial food production due to the lack of processing technologies, difficulties in making the millet flour amenable to the different types of staple foods around the world, for example flatbreads, baked products etc and therefore the consumption is restricted mainly to their growing regions. Recent studies have highlighted the prebiotic properties and beneficial metabolic effects of minor cereals such as millets. Further, increased incidence of diabetes and obesity are driving the RTE food market to functional foods containing complex carbohydrates with higher levels of dietary fibre and health beneficial phytochemicals that can decrease health problems associated with diabetes and obesity. Fortification of diets with food rich in phenolic acids has been shown to have antiglycemic, and antioxidative properties, and can be exploited in developing health foods for everyday consumption.
US2015351415A1 describes millet-based, gluten-free foods and doughs and methods of making them, but it requires the addition of preservatives.
WO2005063026A1 discloses bread made from millet. The bread of
WO2005063026 is different from a flatbread, which is a staple in some parts of the world. Millet cereals are considered a super-food as they are high in nutritional value and are rich in proteins, vitamins, minerals, and fibres. RTE flatbread-based
product based on millet cereals are highly desired. Hence a millet-based flatbread that may overcome some of the shortcomings of existing flatbread RTE product is desirable.
SUMMARY
Embodiments of the present invention provide a ready to eat millet-based flatbread.
One aspect of the invention is a ready to eat millet based flatbread derived from a starter culture wherein the millet based flatbread comprises gluten rich cereal and millet based cereal in a ratio of 1:1 to 2:1 and wherein the starter culture comprises lactic acid bacteria ( LAB) and yeast in a ratio of 3:1 to 5:1.
In one aspect the ready to eat millet based flatbread wherein the flatbread has a sensorial performance comparable to that of a flatbread comprising gluten rich cereal only.
In one aspect the ready to eat millet based flatbread wherein the lactic acid bacteria are Lactobacillus species, Lactobacillus plantarum and Lactobacillus rossiae, Lactobacillus acidophilus, Companilalactobacillus sp., Limosilactobacilluss pontus, Levilactobacillus brevis, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Lactobacillus casei, Bifidobacterium breve, Bifidobacterium infantis, Lactococcus species, Oenococcus species, Streptococcus species, Weissella species, Enterococcus species, Leuconostoc species, Pediococcus species or any combinations thereof.
In one aspect the ready to eat millet based flatbread has a shelf life of up to 4 days at 25-30 ? temperature. In one aspect the ready to eat millet based flatbread has a shelf life of up to 40 days at 2-4? temperature.
One aspect the invention is a method of making a millet-based flatbread of the by mixing a cereal powder comprising a gluten-rich cereal and a millet-based cereal with a starter culture in sufficient amount of water to form a dough, wherein the starter culture comprises lactic acid bacteria and yeast. The method further comprises letting the dough ferment for a time in the range of 2 hours to 24 hours to form a fermented dough.
The method further comprises shaping to form a flatbread-shaped dough after resting the fermented dough. The method further comprises cooking the flatbread shaped dough to form the flatbread.

In another embodiment, a millet-based flatbread is provided. The millet-based flatbread is prepared by a method comprising mixing a cereal powder comprising a gluten-rich cereal and a millet-based cereal with a starter culture in sufficient amount of water to form a dough, wherein the starter culture comprises lactic acid bacteria and yeast. The method further comprises letting the dough ferment for a time in the range of 2 hours to 24 hours to form a fermented dough. The method further comprises shaping to form a flatbread-shaped dough after resting the fermented dough. The method further comprises cooking the flatbread-shaped dough to form the flatbread.

BRIEF DESCRIPTION OF DRAWINGS

The advantages of embodiments of the disclosure may be more readily ascertained from the description of certain examples of the embodiments of the disclosure when read in conjunction with the accompanying drawing, in which:
FIG. 1 is a flowchart 10 of a method of preparing a millet-based flatbread according to embodiments of the disclosure; and
FIG. 2 is a picture of the millet-based flatbreads prepared using the method as illustrated in FIG.1.
FIG. 3 is a flowchart of a method of preparing a millet-based flatbread according to embodiments of the disclosure.
FIG. 4 is a Krona plot showing relative abundance of Lactic acid bacteria (LAB) and yeast species from the SHOTGUN METAGENOME ANALYSIS report.
FIG. 5. Shows TMC, TBC, nutritional attributes, organoleptic properties and storage attributes of flatbread – jowar cooked paratha
FIG. 6. Shows TMC, TBC and nutritional attributes of flatbread –ragi cooked paratha
FIG 7. Shows microbial results of 6 chapati samples at ambient conditions (25-45°C.
FIG 8. Shows microbial results of 6 chapati samples at refrigerated conditions (2-4 °C.

DETAILED DESCRIPTION
The following description and example illustrate some exemplary embodiments of the disclosed invention in detail. Those of skill in the art will recognize that there are numerous variations and modifications of this invention that are encompassed by its scope. Accordingly, the description of a certain exemplary embodiment 25 should not be deemed to limit the scope of the present invention.

The term “comprising” as used herein is synonymous with “including,” or “containing,” and is inclusive or open-ended and does not exclude additional, unrecited elements or method step.
All numbers expressing quantities of ingredients, property measurements, and so forth used in the specification are to be understood as being modified in all instances by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth herein are approximations that may vary depending upon the desired properties sought to be obtained.

Fermentation is an age-old technique whereby chemical breakdown of food occurs due to action of microorganisms resulting in transformation of food.
Typically, the carbohydrates and sugars present in food breaks down to alcohols,
gases or organic acids depending on the type of microorganism. The predominant microorganisms for fermentation of food are lactic acid bacteria (LAB), yeast and certain fungi. It is known that fermentation of cereals breaks down phytates in the grains and makes nutrients available. Further, fermentation can break down gluten thus making the resulting food product more digestible and helps in sugar
management.
The raw food can be fermented by two main methods, namely, “natural fermentation” or “spontaneous fermentation”, or by addition of a “starter culture” or “culture”.
The term “Krona plot” is a a visualization tool designed for metagenomic data exploration. It allows intuitive exploration of relative abundances and confidences within complex hierarchies of metagenomic classifications
The term “starter culture” as used herein, is defined as a preparation of living microorganisms, which are deliberately used to assist the beginning of fermentation, producing specific changes in the chemical composition and the sensorial properties of raw food to obtain a more homogeneous fermented food
product.
“Fermented food product” can be defined as food produced through controlled microbial growth by the enzymatic action on raw food from which they are produced.

In spontaneous fermentation, microorganisms present in the raw food or processing environment breaks down the food. As the microorganisms in spontaneous fermentation are dependent on geographical, and other external factors it brings in a measure of variability and hence in mass production or in large scale production of food it is not preferred. Moreover, natural fermentation is a time consuming process. In the second method of addition of culture, fermentation is initiated by selected commercial starter culture having specific microorganisms. In another variation of addition of starter culture, a small amount of previously fermented batch is added to the raw food and this process of fermentation is known as “back slopping”. The previously fermented batch is otherwise termed as “sourdough culture”. The sourdough culture could be a result of natural fermentation or by addition of a starter culture. Fermented food products are not only rich in nutrients such as proteins, vitamins, essential amino acids, sugars, fatty acids but also good for digestion.

Referring to FIG. 1, FIG. 1 is a flowchart 10 of an exemplary method of preparing a millet-based flatbread in accordance with embodiments of the disclosure. The term “flatbread”, as used herein refers to breads having a thickness ranging from a few millimeters to a few centimeters. Non-limiting examples of flatbreads include pita, phulka, Arabic pita, baladi, mafrood, shami, hapati, sangak, roti, tortillas, naan, taboon, barbari, shrak, mashrouh, nasir, tannoor, lavash, taftan, chapati and pizza.
The flowchart 10 of the method comprises step 12 of mixing a cereal powder comprising a gluten-rich cereal and a millet-based cereal with a starter culture in sufficient amount of water to form a dough.
The gluten-rich cereal comprises wheat; varieties and derivatives of wheat such as maida, wheatberries, durum, emmer, semolina, spelt, farina, farro, graham; rye, barley, and oats. In one preferred embodiment, the gluten-rich cereal is wheat. The gluten-rich cereal is ground using methods known in the art to form part of the cereal powder.
Example millet-based cereal includes pearl millet (bajra) (Pennisetum glaucum); foxtail millet (kangni) (Setaria italica); kodo millet (koden) (Paspalum scrobiculatum); little millet (kutki) (Panicum sumatrense,); barnyard millet (jhangora) (Echinochloa utilis); sorghum (jowar) (Sorghum bicolor L); finger millet (nachani or ragi) (Eleusine coracana); fonio (Digitaria exilis); proso millet (Panicum miliaceum); and tef (Eragrostis tef). In one preferred embodiment, the millet-based cereal is finger millet. The millet-based cereal is ground using methods known in the art to form part of the cereal powder. Alternately, required amounts of the gluten-rich cereal and the millet-based cereal are ground together to form the cereal powder.
One embodiment of the invention is a ready to eat millet based flatbread derived from a starter culture wherein the millet based flatbread comprises gluten rich cereal and millet based cereal in a ratio of 1:1 to 2:1 and wherein the starter culture comprises lactic acid bacteria ( LAB) and yeast in a ratio of 95:1 to 10:1.
In one embodiment the starter culture comprises lactic acid bacteria (LAB) and yeast.in a ratio of 3:1 to 5:1.
In one embodiment the ready to eat millet based flatbread wherein the flatbread has a sensorial performance comparable to that of a flatbread comprising gluten rich cereal only.
In one embodiment the lactic acid bacteria are Lactobacillus species, Lactobacillus plantarum and Lactobacillus rossiae, Lactobacillus acidophilus, Companilalactobacillus sp., Limosilactobacilluss pontus, Levilactobacillus brevis, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Lactobacillus casei, Bifidobacterium breve, Bifidobacterium infantis, Lactococcus species, Oenococcus species, Streptococcus species, Weissella species, Enterococcus species, Leuconostoc species, Pediococcus species or any combinations thereof.
In one embodiment the yeast are Saccharomyces sp, or Saccharomyces cerevisiae.
In one embodiment the flatbread is pita, roti, phulka, Arabic pita, baladi, mafrood, shami, hapati, sangak, tortillas, naan, taboon, barbari, shrak, mashrouh, nasir, tannoor, lavash, taftan, chapati or pizza.
In one embodiment the millet based cereal is derived from pearl millet (bajra) (Pennisetum glaucum); foxtail millet (kangni) (Setaria italica); kodo millet (koden) (Paspalum scrobiculatum); little millet (kutki) (Panicum sumatrense,); barnyard millet (jhangora) (Echinochloa utilis); sorghum (jowar) (Sorghum bicolor L); finger millet (nachani or ragi) (Eleusine coracana); fonio (Digitaria exilis); proso millet (Panicum miliaceum); and tef (Eragrostis tef).
In one embodiment the millet based cereal is derived from pearl millet (bajra)(Pennisetum glaucum); sorghum (jowar) (Sorghum bicolor L) or finger millet (nachani or ragi) (Eleusine coracana). In one embodiment the millet based flatbread does not have any added preservatives.

In one embodiment the millet based flatbread has a moisture content in the range of 28% to 35%. Moisture or humidity content is the ratio of water in grams to flour in gms. This is as measured in the final baked flatbread. In one embodiment the flatbread has a fibre content above 2.7%.
In one embodiment the ready to eat millet based flatbread has a shelf life of up to 4 days at 25-30 ? temperature.
In one embodiment the ready to eat millet based flatbread has a shelf life of up to 40 days at 2-4? temperature.
One embodiment of the invention is a method of making a millet based flatbread derived from a starter culture wherein the millet based flatbread comprises gluten rich cereal and millet based cereal, the method comprising the steps of ;
a. Mixing a cereal powder comprising gluten rich cereal and a millet based cereal in a ratio of 1:1 to 2:1 (wt/wt) with a starter culture comprising LAB and yeast in a ratio of 95:1 to 10:1 along with water to yield a dough
b. Fermenting the dough for a period of 2-24 hours at a temperature of 25-45 ?to yield a fermented dough
c. Shaping the fermented dough to make the flatbread.
In one embodiment water is added at a temperature of 25?-60 ? in step a. In one embodiment the dough is rested for a period of 30-150 minutes at ambient temperature and refrigerated for 8-24 hours at 2-5°C prior to shaping in step c. In one embodiment the dough is fermented at a temperature lower than ambient temperature at step b. In one embodiment the dough comprises additional ingredients selected from seasoning, leavening agent, fats, edible oils, flavour enhancing agents, salt, chilli or a combination thereof. In one embodiment, the starter cultures for the mixing step 12 comprises a mixture of LAB and yeast in a ratio of 5:1. In another embodiment, the starter culture for the mixing step 12 is obtained from a previously fermented batch or “starter culture”. In one embodiment, the previously fermented batch is a result of natural fermentation on cereal powder comprising the gluten-rich cereal alone, or millet based cereal alone, or any combinations thereof. In another embodiment, the starter culture is a result of fermentation by addition of starter culture on the gluten-rich cereal alone, or millet-based cereal alone, or any combinations thereof. The starter culture can be in the form of liquid, semi-liquid or solid. It is known that the microorganisms in starter culture varies depending on parameters such as the types of cereal, water, processing conditions, geographic location, or physical location of the sourdough culture. In one embodiment, the starter culture comprises lactic acid bacteria and yeast.
In one embodiment the millet based flatbread is made from a dough that comprises natural binders (0.2%) like guar gum or xanthan gum. In one embodiment the dough is rested before making the millet based flatbread and comprises LAB to yeast in a ratio between 3:1 to 5:1. (Fig 4)
The term, “lactic acid bacteria (LAB)”, as used herein refers to organism of a group of Gram-positive, catalase negative, non-motile, microaerophilic or anaerobic bacteria which ferment sugars, thereby producing acids including lactic acid as a major product as well as acetic acid, formic acid and/or propionic acid.
The LAB preserve the flatbread so formed by lowering its pH and thus preventing growth of unwanted microorganisms such as mold and thus enhancing shelf-life. Non-limiting examples of LAB includes Lactobacillus species, Lactobacillus plantarum and Lactobacillus rossiae, Lactobacillus acidophilus, Companilalactobacillus sp., Limosilactobacilluss pontus, Levilactobacillus brevis, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Lactobacillus casei, Bifidobacterium breve, Bifidobacterium infantis, Lactococcus species, Oenococcus species, Streptococcus species, Weissella species, Enterococcus species, Leuconostoc species, Pediococcus species or any combinations thereof.
The yeast in the starter culture causes the dough to rise due to release of carbon
dioxide and improves the texture and fluffiness of the millet-based flatbread so formed. Non-limiting example of yeast includes Saccharomyces species, Saccharomyces cerevisiae , or any combinations thereof.
At step 12, (Fig 1) the cereal powder is mixed with the starter culture in sufficient amount of water to form the dough. The dough may additionally include leavening agents. The leavening agents may be added with dry ingredients such as cereal powder. Non-limiting examples of leavening agents include baking soda, baking powder or any combinations thereof.
The dough may additionally include fats and edible oils such as the ones used in cooking. The edible oils and fats may be chosen according to local geographic preference. Non-limiting examples of edible oils and fats include butter, ghee, sunflower oil, safflower oil, groundnut oil, rice bran oil, coconut oil, wheat germ oil, canola oil, olive oil, sesame oil, mustard oil or any combinations thereof.

The water for mixing can be at ambient temperature or at elevated temperature. In one embodiment, the temperature of water is in the range of 25? to 60?. A ratio of the water to the combined amount of the cereal powder and the starter culture is in the range of 1:1.2 to 1:1.6. The mixing may be performed manually or by using mixers known in the art. In one embodiment, the mixing is performed using a stand mixer to form the dough.

At step 14, the dough is let to ferment for a time in the range of 2 hours to 24 hours to form a fermented dough. The fermentation of dough is performed at ambient
temperature, in one embodiment. The ambient temperature is in the range of 25? to 45?. In another embodiment, the dough is let to ferment at ambient temperature followed by maintaining the dough at lower temperature than ambient temperature to form the fermented dough. In one embodiment the lower temperature is in the range of 2-5 °C. This is particularly used for long duration fermentation time beyond 2-4 hrs. In one embodiment the fermentation at ambient temperature range is for 2-4 hours. In one embodiment the fermentation at temperature range lower than ambient temperature range is for up to 24 hours.
At step 16, the fermented dough is shaped to form a flatbread-shaped dough. The “flatbread-shaped dough”, hereinafter is also termed as “shaped dough”. The shape of the flatbread is decided based on the intended flatbread product. For example, if the product is “chapati”, it is preferred the flatbread be round-shaped and if the product is “naan” an oval shape is preferred.
In one embodiment, the fermented dough is brought to ambient temperature and rested prior to the shaping step. The resting of the dough may be carried out for a time in the range of 30 minutes to 2 hours. The shaping of the fermented dough may be performed by hand or by any suitable machines. The shaped dough, in one embodiment, is proofed. The proofing step may additionally help in rising of the shaped dough whereby the fluffiness of the flatbread may be enhanced. The proofing of the dough may be carried out for a time in the range of 30 minutes to 2 hours.
At step 18, the shaped dough is cooked to form the flatbread. The cooking may be performed on a hot cast-iron pan or griddle, or over a stone in a tandoor style oven, or on a stovetop, or in an oven, or a grill. Alternately, the cooking may be performed on industrial-scale hot plates, rollers, pressing machines and the like, such as the ones used for pizza or tortilla making.
The cooking, in one embodiment, is performed at temperatures in excess of 180? and up to a temperature of 500?.
The flatbread of the present disclosure can contain, for example, cheese, herbs and spices, fruits, vegetables, encapsulated flavour and/or aroma ingredients, and the like as well as mixtures thereof. Additionally, the flatbread of the present disclosure can contain topical applications of, or be coated with, ingredients such as, for
example, sunflower seeds, sesame seeds, pumpkin seeds, chia seeds, spice blends and the like as well as mixtures thereof.

The flatbreads formed according to embodiments of the present disclosure is packed in a pouch, tray, envelope, or the like, under vacuum, or inert gas and/or inert gas flushed conditions (e.g., low oxygen and low moisture atmosphere). In certain embodiments, the packaged flatbread may be frozen to enhance shelf-life.
FIG.2 includes pictures of various flatbreads formed according to the method as described in FIG.1. A particular advantage of the inventive flatbread as compared
to existing millet-based flatbread is its longer shelf-life due to anti-mold effect due to presence of the starter culture. The shelf-life of the millet-based flatbread is up to two weeks when stored in ambient temperature. The shelf-life can be prolonged for up to six months when maintained under refrigeration at temperatures below 5?. The flatbread of the present disclosure has increased organoleptic properties
such as taste, softness, fluffiness and texture when compared to existing millet based flatbread.

EXAMPLES
Example 1: Comparative Example
Preparation of control samples: About 250 grams of whole wheat flour was taken in a container. About 3 grams of salt and 0.9 grams of baking soda were added to the container and mixed. About 30 grams of starter culture was added to the container along with 20 grams of oil. About 185 ml of filtered water was added to the container and mixed to form the dough and was left to ferment for 2 hours. The fermented dough was then kept in the refrigerator overnight for further fermentation. The fermented dough was brought to room temperature and shaped
and cooked over a pan to form wheat chapati. The amount of starter culture and baking soda were optimised to get best results.

Similarly, a refined wheat flour-based naan was prepared. The naan was prepared from 250 grams of refined wheat flour, 50 grams of yoghurt, 5 grams of sugar, 5 grams of salt, 100 ml of warm milk, 1.25 grams of baking soda, 0.5 grams of baking powder, 10 ml oil and 10 ml of water, in a manner similar to the preparation of wheat chapati. The wheat chapati and naan were analysed for microbial activity and sensorial performance. The microbial activity expressed as Total Bacteria Content (TBC) was recorded at ambient temperature (25?) and at refrigerated condition (at 4?) over many days. The wheat chapati and naan were found to have minimal TBC at ambient temperature for two days and under refrigerated condition they remained microbe-free for even 21 days. With respect to sensorial performance as to consumer acceptance in terms of appearance, colour, texture, taste and flavour, the wheat chapati and naan were evaluated over time at ambient temperature and at refrigerated storage condition, varying amounts of starter culture, and time for fermentation. With overnight fermentation sensorial performance improved and the samples remained within specification at ambient temperature for two days and under refrigerated condition for 20 days.

Example 2: Preparation of Chapatis
Preparation of millet-based flatbreads: About 150 grams of whole wheat flour was taken with 100 grams of finger millet (ragi) flour in a container. About 3 grams of salt and 0.9 grams of baking soda were added to the container and mixed. 30 grams of starter culture was added to the container along with 20 grams of oil. About 185 ml of filtered water was added to the container and mixed to form the dough and was left to ferment for 2 hours. The fermented dough was then kept in the refrigerator overnight for further fermentation. The fermented dough was brought to room temperature and shaped and cooked over a pan to form ragi chapati. Similarly, flatbreads were prepared from sorghum (jowar), and pearl millet (bajra) flour to form jowar chapati and bajra chapati, respectively.

EXAMPLE 3 – AMOUNT OF STARTER

Methodology
Starter development: Where 1:1 ratio of wheat flour and water were used and feed was given for every 12 hours for 3 days and was used in the product development. To optimize the usage of starter quantity, it was varied from 20gms to 50gms to prepare 3 different product samples of each flatbread type

For standardizing the amount of starter

NAAN
S.No INGREDIENTS (in grams) QUANTITY (in grams)
1 Maida 250 250 250
2 Yoghurt 50 50 50
3 Starter 30 40 50
4 Warm milk 100 100 100
5 Salt 5 5 5
6 Sugar 5 5 5
7 Soda 1.25 1.25 1.25
8 Water 10 10 10
9 Oil 10 10 10
10 Baking powder 0.5 0.5 0.5

WHEAT CHAPATI
S.No INGREDIENTS QUANTITY (in grams)
1 Flour 250 250 250
2 Starter 20 30 40
3 Oil 15 15 15
4 Soda 0.9 0.9 0.9
5 Salt 3 3 3
6 Filtered water 185 185 185

BAJRA/RAGI/JOWAR CHAPATIS
S.No INGREDIENTS QUANTITY
1 Wheat flour 150 150 150
2 Bajra/ragi/jowar 100 100 100
3 Starter 20 30 40
4 Oil 15 15 15
5 Soda 0.5 0.9 1.5
6 Salt 3 3 3
7 Filtered water 185 (180 bajra) 185 (180 bajra) 185 (180 bajra)

WHEAT PHULKA
S.No INGREDIENTS QUANTITY
1 Flour 250 250 250
2 Starter 20 30 40
3 Oil 10 10 10
4 Soda 0.9 0.9 0.9
5 Salt 3 3 3
6 Filtered water 185 185 185

Moisture Content and Fibre analysis
The millet-based chapatis were found to have a moisture content in the range of 29.47% to 31.8%7 as compared to 29.79% for wheat chapati. The fibre content of these chapatis were in the range of 2.76% to 3.23% as compared to 2.58% for wheat chapati.
The chapatis were tested for microbial activity to evaluate the shelf-life. At ambient room temperature, the millet-based chapatis had a shelf-life of 3 days and when stored under refrigerated condition it was found to be more than 30 days.

EXAMPLE 4 – SENSORY ANALYSIS
Sensory evaluation: The sensory evaluation was done for the prepared samples, using 9-point hedonic scale.
Nutritional assessment: The best accepted samples were selected for nutritional analysis i.e., proximate analysis (moisture, protein, fat, ash, carbohydrates, crude fiber, energy).
The sensorial performance of millet-based flatbreads was evaluated and they were found to be at par with the wheat chapati and naan. The sensorial performance of millet-based flatbreads was found to improve on storage for 1 day at ambient temperature and their performance were much better than wheat chapati.
The sensory evaluation was done for the prepared samples, using 9-point hedonic scale.

Tables 1-12 show data on sensory analysis with and without refrigeration
Proportion 1 : 20 grams of starter culture
Proportion 2 : 30 grams of starter culture
Proportion 3 : 40 grams of starter culture
Table 1
Wheat chapati without refrigeration
Samples Apperance Colour Texture Taste Flavour Overall acceptability
Proportion 1 8.33 8.26 8.13 8.2 8.2 8.06
Proportion 2 9 9 8.73 8.73 8.6 8.93
Proportion 3 8.62 8.18 7.75 8.37 8.12 8.31

Table 2
Bajra without Refrigration
Samples Apperance Colour Texture Taste Flavour Overall acceptability
Proportion 1 7.93 8.06 7.73 8.06 7.8 7.93
Proportion 2 8.86 8.86 8.73 8.53 8.4 8.8
Proportion 3 8.56 8.06 7.8 7.87 8.06 8.25

Table 3
Phulka without Refrigration
Samples Apperance Colour Texture Taste Flavour Overall acceptability
Proportion 1 7.93 8 7.6 7.53 7.66 7.73
Proportion 2 8.13 7.8 7.6 7.66 7.53 7.86
Proportion 3 8.13 8 7.86 7.66 7.46 7.66

Table 4
Jowar without Refrigeration
Samples Appearance Colour Texture Taste Flavour Overall acceptability
Proportion 1 8.26 8.13 8 7.93 7.86 7.93
Proportion 2 8.66 8.53 8.53 8.4 8.26 8.46
Proportion 3 8.37 8 7.87 7.93 7.93 7.93

Table 5
Ragi chapati without Refrigeration
Samples Appearance Colour Texture Taste Flavour Overall acceptability
Proportion 1 7.6 7.73 8 7.86 7.8 7.8
Proportion 2 8.26 7.86 8.06 7.86 8.26 8
Proportion 3 8.25 7.81 7.75 7.62 8 7.62
Table 6
Naan without Refrigeration
Samples Appearance Colour Texture Taste Flavour Overall acceptability
Proportion 1 8.26 8.13 7.73 7.93 7.93 7.93
Proportion 2 8.33 8.26 8 8.2 8.13 8.2
Proportion 3 8.13 8 7.93 7.8 7.86 7.93

With Refrigeration:

Table 7
Wheat chapati with refrigration
Samples Apperance Colour Texture Taste Flavour Overall acceptability
Proportion 1 8.75 8.56 8.81 8.62 8.62 8.62
Proportion 2 8.29 8.17 8.35 8.05 8.58 8.41
Proportion 3 8.06 7.81 7.81 7.68 7.81 7.81

Table 8
Bajra chapati with refrigration
Samples Apperance Colour Texture Taste Flavour Overall acceptability
Proportion 1 8.18 8.25 7.87 8 7.87 7.93
Proportion 2 8.05 8.29 8.05 8.17 8.17 8.35
Proportion 3 8.31 8.12 8.06 7.93 7.93 8

Table 9
Phulka with refrigeration
Samples Appearance Colour Texture Taste Flavour Overall acceptability
Proportion 1 8.37 8.31 8.37 8.18 8.18 8.43
Proportion 2 8.11 7.94 7.94 8.05 8.11 8.23
Proportion 3 8.11 7.94 7.94 8.05 8.11 8.23

Table 10
Jowar chapati with refrigeration
Samples Appearance Colour Texture Taste Flavour Overall acceptability
Proportion 1 8.56 8.43 8.37 8.12 8.25 8.12
Proportion 2 8.23 8.17 7.88 7.94 8.11 8.05
Proportion 3 8.75 8.56 8.56 8.31 8.43 8.62

Table 11
Ragi chapati with refrigeration
Samples Appearance Colour Texture Taste Flavour Overall acceptability
Proportion 1 8.75 8.18 8.25 7.8125 7.87 8
Proportion 2 8.29 8.05 8 8.11 8.47 8.17
Proportion 3 7.87 7.68 8.18 7.81 7.93 7.75

Table 12
Naan chapati with refrigration
Samples Apperance Colour Texture Taste Flavour Overall acceptability
Proportion 1 8.18 7.81 7.56 7.56 7.56 7.56
Proportion 2 7.76 7.88 7.52 7.64 7.64 7.82
Proportion 3 7.75 7.5 7.5 7.43 7.43 7.43

Proportion 2 was best for both overnight refrigeration and two hours fermentation for all the six samples but the rolling properties for the dough was good for the overnight refrigerated samples.
In this combination of Proportion 2, the Millet based chapatis are compared to standard wheat chapati for different sensory properties and overall acceptability.
Table 13
Sample Appearance Color Texture Taste Flavour Overall acceptability
Wheat chapati (C21) 8.29 8.17 8.35 8.05 8.58 8.41
Phulka (C25) 8.11 7.94 7.94 8.05 8.11 8.23
Ragi chapati (C22) 8.29 8.05 8 8.11 8.47 8.17
Bajra chapati (C24) 8.05 8.29 8.05 8.17 8.17 8.35
Jowar chapati (C23) 8.23 8.17 7.88 7.94 8.11 8.05
Naan (C26) 7.76 7.88 7.52 7.64 7.64 7.82

EXAMPLE 5 – STORAGE STUDIES
Shelf life studies: The best accepted samples were stored for 1, 3, 5, 7, 8, 9, 11 till 21 days and were evaluated for the microbial quality (both bacterial and mold)
Packaging material: The best accepted samples were stored in PET PE, Thick LDPE and Aluminium packaging material.
Storages temperatures: The selected samples were stored at ambient temperatures (25ºC) and refrigerated temperatures 4ºC for shelf life studies.The outcome of shelf life studies is depicted in Figures 7 and 8 pertaining to different flatbread samples. The results are shown for refrigerated and ambient temperature conditions.
The results show that TBC and TMC values in Fig 7 (ambient conditions) for millet based flatbreads are equivalent to that of only gluten rich cereal based flatbreads.
Figure 5 and 6 show results of microbial count for flatbreads of Jowar and Ragi cooked paratha samples.

Example 6 : SHOTGUN METAGENOME ANALYSIS REPORT OF DOUGH
Objectives : Data Quality Control, Metagenome Assembly with MEGAHIT , Binning with CONCOCT, metaBAT2 Checking of bins with CheckM , Taxonomic assignment and functional annotation of bins
METHODOLOGY 1. Raw reads of samples were subjected to adapter trimming with fastp and quality of adapter trimmed reads were checked using FastQC. 2. Metagenome assembly was performed on clean reads of all samples using MEGAHIT assembler. 3. Metagenome Assembled Genomes (MAGs) were extracted from the assembly by a process called ‘binning’ for all samples. 4. The bins extracted from the assemblies were refined further to obtain MAGs which are complete in terms of the completion with lesser contamination. 5. Taxonomic classification was performed for the reconstructed MAGs using taxonomy databases from NCBI. 6. Functional annotation of MAGs was performed using, which locates and translates the ORF regions on MAG contigs into protein sequences which can be further used for downstream analysis.
Table 15: Taxonomic classification of Bins

Software List

Figure 4 shows the Krona plot and the relative abundance of lactic acid bacteria and yeast. In fermented dough rested for 24 hours. The ratio of LAB to yeast is between 3:1 to 5:1
It is to be understood that the above description is intended to be illustrative, and not restrictive. Furthermore, many other embodiments will be apparent to those of skill in the art upon reading and understanding the above description. Although the present invention has been described with reference to specific exemplary 30 embodiments, it will be recognized that the invention is not limited to the embodiments described, but can be practiced with modification and alteration within the spirit and scope of the appended claims.

(Puneeta Arora) IN/PA – 1648
Agent for the applicant
19th day of May 2024
,CLAIMS:1. A ready to eat millet based flatbread derived from a starter culture wherein the millet based flatbread comprises gluten rich cereal and millet based cereal in a ratio of 1:1 to 2:1 and wherein the starter culture comprises lactic acid bacteria ( LAB) and yeast.in a ratio of 95:1 to 10:1.
2. The ready to eat millet based flatbread wherein the flatbread has a sensorial performance comparable to that of a flatbread comprising gluten rich cereal only.
3. The ready to eat millet based flatbread of claim 1 wherein the lactic acid bacteria are Lactobacillus species, Lactobacillus plantarum and Lactobacillus rossiae, Lactobacillus acidophilus, Companilalactobacillus sp., Limosilactobacilluss pontus, Levilactobacillus brevis, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Lactobacillus casei, Bifidobacterium breve, Bifidobacterium infantis, Lactococcus species, Oenococcus species, Streptococcus species, Weissella species, Enterococcus species, Leuconostoc species, Pediococcus species or any combinations thereof.
4. The ready to eat millet based flatbread of claim 1 wherein the flatbread is pita, roti, phulka, Arabic pita, baladi, mafrood, shami, hapati, sangak, tortillas, naan, taboon, barbari, shrak, mashrouh, nasir, tannoor, lavash, taftan, chapati or pizza.
5. The ready to eat millet based flatbread of claim 1 wherein the millet based cereal is derived from millet-based cereal includes pearl millet (bajra) (Pennisetum glaucum); foxtail millet (kangni) (Setaria italica); kodo millet (koden) (Paspalum scrobiculatum); little millet (kutki) (Panicum sumatrense,); barnyard millet (jhangora) (Echinochloa utilis); sorghum (jowar) (Sorghum bicolor L); finger millet (nachani or ragi) (Eleusine coracana); fonio (Digitaria exilis); proso millet (Panicum miliaceum); and tef (Eragrostis tef).
6. The ready to eat millet based flatbread of claim 1 wherein the flatbread has a moisture content in the range of 28% to 35%.
7. The ready to eat millet based flatbread of claim 1 wherein the flatbread has a fibre content above 2.7%.
8. The ready to eat millet based flatbread of claim 1 with a shelf life of upto 4 days at 25-30 ? temperature.
9. The ready to eat millet based flatbread of claim 1 with a shelf life of up to 45 days at 2-4? temperature.
10. A method of making a millet based flatbread derived from a starter culture wherein the millet based flatbread comprises gluten rich cereal and millet based cereal, the method comprising the steps of ;
a. Mixing a cereal powder comprising gluten rich cereal and a millet based cereal in a ratio of 1:1 to 2:1 (wt/wt) with a starter culture comprising LAB and yeast in a ratio of 95:1 to 10:1 along with water to yield a dough
b. Fermenting the dough for a period of 2-24 hours at a temperature of 25-45 ?to yield a fermented dough
c. Shaping the fermented dough to make the flatbread.
11. The method of claim 10 wherein the water is added at a temperature of 25?-60 ? in step a.
12. The method of claim 10 wherein the dough is rested for a period of 30-150 minutes at ambient temperature and refrigerated for 8-24 hours at 2-5 Deg C prior to shaping in step c.
13. The method of claim 10 wherein the dough is fermented at a temperature lower than ambient temperature at step b.
14. The method of claim 10 wherein the dough comprises additional ingredients selected from seasoning, leavening agent, fats, edible oils, flavour enhancing agents, salt, chilli or a combination thereof.